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Lentiviral Mediated Gene Silencing in Human Pseudoislet Prepared in Low Attachment Plates

Article DOI: 10.3791/59578-v • 11:24 min • May 14th, 2019
May 14th, 2019 •
1,2, 1,2, 1,2, 2,3, 2,3, 1,2
1Department of Internal Medicine, Carver College of Medicine, University of Iowa, 2Fraternal Order of Eagles Diabetes Research Center, University of Iowa, 3Roy J. Carver Department of Biomedical Engineering, University of Iowa

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Summary
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A protocol to create gene modified human pseudoislets from dispersed human islet cells that are transduced by lentivirus carrying short hairpin RNA (shRNA) is presented. This protocol utilizes readily available enzyme and culture vessels, can be performed easily, and produces genetically modified human pseudoislets suitable for functional and morphological studies.

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Gene Silencing Lentiviral Mediated Gene Silencing Human Islets Low Attachment Plates Lentivirus Mediated SHR8 Transduction Islet Function Downregulation Of Gene Expression Prolonged Culture Suspension Shipping Medium 50 Milliliter Conical Centrifuge Tube Biological Safety Cabinet Islet Pellet CMRL Medium Human Serum Albumin Non-tissue Culture Treated Dish Overnight Culture 15 Milliliter Conical Centrifuge Tube Swinging Bucket Rotor PBS
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